EMBO workshop 2021

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The tutorials listed here take place virtually on the 11th and 13th of September of 2021 (14:00-18:30) as part of the EMBO workshop for Image processing in cryoEM.

Connect to Guacamole and run Dynamo

Credentials

Please refer to your personal course material to access your two sets of credentials:

  1. A set of credentials to enter Guacamole (the platform for connecting via browser). These are common to all participants.
  2. A set of course credentials to enter the actual machines on which the course will be done: the EMBO21 UserID. These are unique to each participant.

Connect to Guacamole

To connect to Guacamole, first go to the following link in your browser:

https://login.cryst.bbk.ac.uk/guacamole/#/ 

At this stage you should maximize your browser window, otherwise you might have visualization problems when opening Dynamo. You should see the following window. Enter your Guacamole credentials.

Entering guacamole server.

Connect to your machine

Now you need to select a machine. Please then click on +em and then +Servers - Public and select server Zeus-0-14.

Select a machine.

When you select the machine, you will prompted to use your EMBO21 UserID

Entering your personal course credentials.

This should access your virtual Linux XFCe desktop, which should look like this in your browser

Linux XFCe desktop

If you open a terminal (with the terminal icon) you can test by typing

pwd

that you are in the correct path, which should be:

 /d/embo2021/u/emboXX

where XX stands for your actual workshop participant number.

Load and run Dynamo

First you load the standalone version of Dynamo:

 module load dynamo/v1.1.524

and then it can be run by simply typing:

 dynamo
Dynamo console started.

Dynamo specific commands are then typed into this so called Dynamo console. For example, try to type the Dynamo command

dynamo_version

to see the current version that you are using. Note: Copy paste of commands from your local desktop into the dynamo standalone console on the remote Linux XFCe desktop might not always work. We suggest to additionally open the tutorials that you will be working on in a browser inside the remote Linux XFCe desktop itself, in case you need to copy paste long commands.

Data location

The data used in the tutorials is already available for you. Move to the directory of the corresponding practical using the command:

cd prac-5

or

cd prac-6

respectively. Double-check if the data is present by typing

ls

You should see the following files:

files

Time schedule

The course is divided into different tutorials, described in the time schedule below. Each tutorial is structured in three stages:

  1. Demostration: All students are connected to the main room. The goals of the tutorial will be explained by an instructor.
  2. Individual work: Students go the assigned breakout rooms to work on the tutorials independently. An instructor will be assigned to each breakout room to help with questions.
  3. Common discussion: In the last 5 minutes of the tutorial, we come back to the general breakout room to wrap up the module.

This time schedule is tentative

Day 1: Tomography

Time Tutorial
14:00 Introduction
14:30 Tilt series alignment: manually
15:00 Tilt series alignment: GUI
15:30 Tilt series alignment: command line
16:00 Break
16:30 Models: catalogue system
17:00 Models: helical symmetry
17:30 Models: vesicles and surfaces
18:00 Template matching


Day 2: Subtomogram averaging

Time Tutorial
14:00 Introduction
14:30 Getting started: synthetic thermosomes
15:30 Advanced tutorial: FHV
16:00 Break
16:30 FHV: continuation
17:00 Seed oversampling
18:00 High resolution pipelines
18:20 Closing remarks

Turorials

Alignment of tilt series

We will align the same tilt series tilt series with two different approaches. We prepared the first tilt series from the EMPIAR entry 10164, depicting a set of virus like particles (VLP). Move to the directory that contains this data using the command:

cd prac-5

Double-check by typing

ls

to see all files in the directory. One file should be the following tilt series:

b001ts001.mrc

This is a binned version (for quicker processing) with the pixel size is 2.7 Angstrom (original deposition is 1.35).

Automated alignment of tilt series through the GUI

Walkthrough on GUI based tilt series alignment

Automated alignment of tilt series through the command line

Walkthrough on command line based tilt series alignment

Models

Catalogues

Helical models

tutorial

Vesicle and surface models

tutorial on membrane modeling with dmslice

Reusing models

Reusing model workflows ( walkthrough)

Template matching

walkthrough for automated identification of proteosomes on a real tomogram through template matching. (~1 hour)
The data is available at:

/d/embo2021/d/dynamo/t20s.mrc

An example of template and its mask are provided as 'average32.em' and 'maskTight32.em'.

Subtomogram averaging

Introduction

Guided presentation:

Working on your own:

Basic walkthrough: creating a catalogue, picking particles, launching a project.

Advanced starters guide

Complete the advanced starters guide (~2 hours) The data can be found in

/d/embo2021/d/dynamo/crop.rec 

To use chimera path you need in the tutorial is

 TBI 

Seed oversampling

Tutorial from densely packed spherical geometry (~1 hour). The data is available at:

/d/embo2021/d/dynamo/v17.rec

Since the final alignment project is run on the standalone version of Dynamo, the 'destination' parameter should be 'standalone' instead of 'matlab_parfor'

High resolution pipelines

VLPs

Exercise to train a realistic case including all processing steps from catalogue creation to final structure using a reduced dataset. Start with the exercise and refer to the complete walkthrough for guidance or the solution.

Interconnectivity tools

Alister: Not a tutorial per se: this is a presentation on a set of tools to interconnect Dynamo, M and other software.

A set of tools have been developed which provide a means to use the geometrical tools and alignment projects from Dynamo with data preprocessed in Warp. These tools mean that particle picking and alignment can be performed in Dynamo prior to multi-particle refinement in M.